DOCSLIB.ORG

Selective Effects of PDE10A Inhibitors on Striatopallidal Neurons Require Phosphatase Inhibition by DARPP-321,2,3

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Selective Effects of PDE10A Inhibitors on Striatopallidal Neurons Require Phosphatase Inhibition by DARPP-321,2,3 New Research Disorders of the Nervous System Selective Effects of PDE10A Inhibitors on Striatopallidal Neurons Require Phosphatase Inhibition by DARPP-321,2,3 Marina Polito,1,2 Elvire Guiot,1,2 Giuseppe Gangarossa,3,4,5 Sophie Longueville,2,6,7 Mohamed Doulazmi,1,2 Emmanuel Valjent,3,4 Denis Hervé,2,6,7 Jean-Antoine Girault,2,6,7 Danièle Paupardin-Tritsch,1,2 Liliana R. V. Castro,1,2 and Pierre Vincent1,2 DOI:http://dx.doi.org/10.1523/ENEURO.0060-15.2015 1CNRS, UMR8256 “Biological Adaptation and Ageing”, Institut de Biologie Paris-Seine (IBPS), F-75005 Paris, France, 2Université Pierre et Marie Curie (UPMC, Paris 6), Sorbonne Universités, Paris, F-75005, France, 3CNRS, UMR-5203, Institut de Génomique Fonctionnelle, Montpellier, F-34094, France, 4Institut National de la Santé et de la Recherche Médicale, U661, Montpellier, F-34094, France, 5Universités de Montpellier 1 & 2, UMR-5203, Montpellier, F-34094, France, 6Institut National de la Santé et de la Recherche Médicale UMR-S 839, Paris, France, and 7Institut du Fer a` Moulin, Paris, France Abstract Type 10A phosphodiesterase (PDE10A) is highly expressed in the striatum, in striatonigral and striatopallidal medium- sized spiny neurons (MSNs), which express D1 and D2 dopamine receptors, respectively. PDE10A inhibitors have pharmacological and behavioral effects suggesting an antipsychotic profile, but the cellular bases of these effects are unclear. We analyzed the effects of PDE10A inhibition in vivo by immunohistochemistry, and imaged cAMP, cAMP- dependent protein kinase A (PKA), and cGMP signals with biosensors in mouse brain slices. PDE10A inhibition in mouse striatal slices produced a steady-state increase in intracellular cAMP concentration in D1 and D2 MSNs, demonstrating that PDE10A regulates basal cAMP levels. Surprisingly, the PKA-dependent AKAR3 phosphorylation signal was strong in D2 MSNs, whereas D1 MSNs remained unresponsive. This effect was also observed in adult mice in vivo since PDE10A inhibition increased phospho-histone H3 immunoreactivity selectively in D2 MSNs in the dorsomedial striatum. The PKA-dependent effects in D2 MSNs were prevented in brain slices and in vivo by mutation of the PKA-regulated phosphorylation site of 32 kDa dopamine- and cAMP-regulated phosphoprotein (DARPP-32), which is required for protein phosphatase-1 inhibition. These data highlight differences in the integration of the cAMP signal in D1 and D2 MSNs, resulting from stronger inhibition of protein phosphatase-1 by DARPP-32 in D2 MSNs than in D1 MSNs. This study shows that PDE10A inhibitors share with antipsychotic medications the property of activating preferentially PKA-dependent signaling in D2 MSNs. Key words: biosensor imaging; cAMP; phosphodiesterase; protein kinase; schizophrenia; striatum Significance Statement The striatum is mainly composed of medium-sized spiny neurons that express either dopamine D1 receptors or dopamine D2 receptors. Their activity is associated with either the initiation of movement or action suppression, respectively. Biosensor imaging revealed that pharmacological inhibition of type 10A phosphodiesterase in- creased cAMP levels in D1 and D2 neurons in the same manner, but only D2 neurons exhibited an increase in the protein kinase A-mediated phosphorylation level. This effect resulted from an asymmetrical regulation of phosphatases by DARPP-32. D2 neurons are thus more prone to respond to a tonic cAMP signal than D1 neurons, a property that may explain how phosphodiesterase 10A inhibitors produced antipsychotic-like behavioral effects. This D1/D2 imbalance may also be critical for reward-mediated learning and action selection. July/August 2015, 2(4) e0060-15.2015 1–15 New Research 2 of 15 Introduction Classical and atypical antipsychotic agents share the Schizophrenia is a devastating psychiatric disease, property of inhibiting D2 receptors and thus, in the stria- which results in persistent cognitive and emotional impair- tum, increase PKA-dependent phosphorylation selec- ments. Type 10A phosphodiesterase (PDE10A) inhibitors tively in D2 MSNs (Bateup et al., 2008; Bertran-Gonzalez were recently proposed as a treatment for schizophrenia et al., 2008, 2009). In contrast, psychostimulants, which (Kehler and Nielsen, 2011; Chappie et al., 2012); however, are psychotomimetic, activate many signaling responses their cellular mechanisms of action remain unclear with in D1 MSNs (Bateup et al., 2008; Bertran-Gonzalez et al., respect to their putative therapeutic effects. PDE10A is 2008, 2009). PDE10A inhibitors were shown to increase highly and almost exclusively expressed in medium-sized cAMP levels in the striatum (Schmidt et al., 2008) and spiny neurons (MSNs) of the striatum (Seeger et al., 2003; could be expected to mimic the effects of both antipsy- Coskran et al., 2006; Heiman et al., 2008; Lakics et al., chotic and psychotomimetic compounds. We used bio- 2010; Kelly et al., 2014). MSNs are divided into two cat- sensor imaging to precisely analyze the effects of PDE10A egories based on their expression of dopamine receptors inhibitors on cAMP/PKA signaling at the level of individual and their sites of projection, as follows: MSNs projecting D1 and D2 MSNs. Our work revealed that although PDE10A inhibition increased intracellular cAMP levels in to the substantia nigra highly express dopamine D1 re- both D and D MSNs, the downstream consequences at ceptors (hereafter termed D1 MSNs); whereas, MSNs pro- 1 2 jecting to the external globus pallidus highly express the level of PKA targets were profoundly different: the cAMP signal resulting from PDE10A inhibition strongly adenosine A2A and dopamine D2 receptors (hereafter increased PKA-dependent phosphorylation in D MSNs, termed D2 MSNs; Gerfen et al., 1990; Le Moine and Bloch, 2 1995; Bertran-Gonzalez et al., 2008; Matamales et al., whereas D1 MSNs remained mostly unaffected. Further 2009). The high expression of PDE10A in MSNs, and its analyses showed that the difference required DARPP-32- interaction with the scaffold protein AKAP150 (A-kinase dependent regulation of phosphatase activity in D1 and D2 anchoring protein 150), protein kinase A (PKA), PSD-95, MSNs. and NMDA receptor suggests an important role in mod- ulating the spread of the synaptic cAMP signals into the Materials and Methods cell body (Russwurm et al., 2015). Animals Besides PDE10A, striatal neurons express a number of Animals were housed under standardized conditions specific signaling proteins that markedly differ from those in with a 12 h light/dark cycle, stable temperature (22 Ϯ other brain regions (Girault, 2012), and that determine the 1ºC), controlled humidity (55 Ϯ 10%), and food and water characteristics of the cAMP/PKA signaling pathway (Castro available ad libitum. Homozygous mice expressing et al., 2013). Among these specific proteins, DARPP-32 DARPP-32 with the T34A or T75A mutation (Svenning- (32-kDa dopamine and cAMP-regulated phosphoprotein) is sson et al., 2003) were obtained by crossing heterozygous a multifunctional protein regulating phosphatase and kinase mice, on a mixed C57BL6/J-Sv129 background (a gift of activities: for example, when DARPP-32 is phosphorylated Dr. P. Greengard, The Rockefeller University, New York). at threonine 34 residue (Thr34) by PKA, it becomes a potent Male Drd2-EGFP heterozygous mice (C57Bl6/J) were inhibitor of serine/threonine protein phosphatase-1 (PP-1; generated as described previously (Gong et al., 2003). Hemmings et al., 1984; Svenningsson et al., 2004), increas- Experiments were performed in accordance with the reg- ing the duration of PKA-dependent signals (Castro et al., ulations under the control of the local ethic committee 2013). Charles Darwin C2EA - 05. Live brain slice preparation Received June 1, 2015; accepted August 10, 2015; First published August 25, 2015. Brain slices were prepared from male mice that were 1The authors declare no competing financial interests. 8–12 days of age. Coronal brain slices were cut with a 2Author contributions: M.P., E.V., D.H., D.P.-T., L.R.V.C., and P.V. designed VT1200S microtome. Slices were prepared in an ice-cold research. M.P., E.G., G.G., S.L., and L.R.V.C. performed research. M.P., E.G., solution of the following composition (in mM): 125 NaCl, 0.4 G.G., S.L., M.D., L.R.V.C., and P.V. analyzed data. E.V., D.H., J.A.-G., D.P.-T., CaCl , 1 MgCl , 1.25 NaH PO , 26 NaHCO , 25 glucose, L.R.V.C., and P.V. wrote the paper. 2 2 2 4 3 3This work was supported by grants from ATIP-Avenir (Inserm) and from the and 1 kynurenic acid, saturated with 5% CO2 and 95% O2. Agence Nationale de la Recherche, ANR-2010-JCJC-1412) to EV and ANR09- The slices were incubated in this solution for 30 min and then MNPS-014 to DH, and ERC to JAG. The groups of PV, and JAG and DH are placed on a Millicell-CM membrane (Millipore) in culture part of the Bio-Psy Laboratory of Excellence. medium (50% Minimum Essential Medium, 50% HBSS, 6.5 Acknowledgments: Confocal microscopy and image analysis were per- formed at the Institut du Fer a` Moulin Imaging Facilities and at the Institute of g/L glucose, penicillin-streptomycin; Invitrogen). We used Biology Paris-Seine Imaging Facility (supported by the “Conseil Regional Ile-de the Sindbis virus as a vector to induce expression of the France”, the French National Research Council, and Sorbonne University, various biosensors after overnight incubation (Ehrengruber UPMC, Paris 6). et al., 1999). The coding sequences of Epac-SH150 (Polito Correspondence should be addressed to Pierre Vincent, UMR8256, 9, quai et al., 2013), AKAR2-NLS (Zhang et al., 2005), AKAR3 (Allen St. Bernard, F-75005 PARIS, France. E-mail: [email protected]. DOI:http://dx.doi.org/10.1523/ENEURO.0060-15.2015 and Zhang, 2006), and cygnet2 (Honda et al., 2001) were Copyright © 2015 Polito et al. inserted into the viral vector pSinRep5 (Invitrogen). The viral This is an open-access article distributed under the terms of the Creative vector (ϳ5 ϫ 105 particles per slice) was added, and slices Commons Attribution 4.0 International, which permits unrestricted use, distri- bution and reproduction in any medium provided that the original work is were incubated overnight at 35°C under an atmosphere properly attributed.
Load more

Recommended publications
  • Structure-Based Redesigning of Pentoxifylline Analogs Against
    www.nature.com/scientificreports OPEN Structure‑based redesigning of pentoxifylline analogs against selective phosphodiesterases to modulate sperm functional competence for assisted reproductive technologies Mutyala Satish1,5, Sandhya Kumari2,5, Waghela Deeksha1, Suman Abhishek1, Kulhar Nitin1, Satish Kumar Adiga2, Padmaraj Hegde3, Jagadeesh Prasad Dasappa4, Guruprasad Kalthur2* & Eerappa Rajakumara1* Phosphodiesterase (PDE) inhibitors, such as pentoxifylline (PTX), are used as pharmacological agents to enhance sperm motility in assisted reproductive technology (ART), mainly to aid the selection of viable sperm in asthenozoospermic ejaculates and testicular spermatozoa, prior to intracytoplasmic sperm injection (ICSI). However, PTX is reported to induce premature acrosome reaction (AR) and, exert toxic efects on oocyte function and early embryo development. Additionally, in vitro binding studies as well as computational binding free energy (ΔGbind) suggest that PTX exhibits weak binding to sperm PDEs, indicating room for improvement. Aiming to reduce the adverse efects and to enhance the sperm motility, we designed and studied PTX analogues. Using structure‑guided in silico approach and by considering the physico‑chemical properties of the binding pocket of the PDEs, designed analogues of PTX. In silico assessments indicated that PTX analogues bind more tightly to PDEs and form stable complexes. Particularly, ex vivo evaluation of sperm treated with one of the PTX analogues (PTXm‑1), showed comparable benefcial efect at much lower concentration—slower
    [Show full text]
  • Phosphodiesterase 1B Knock-Out Mice Exhibit Exaggerated Locomotor Hyperactivity and DARPP-32 Phosphorylation in Response to Dopa
    The Journal of Neuroscience, June 15, 2002, 22(12):5188–5197 Phosphodiesterase 1B Knock-Out Mice Exhibit Exaggerated Locomotor Hyperactivity and DARPP-32 Phosphorylation in Response to Dopamine Agonists and Display Impaired Spatial Learning Tracy M. Reed,1,3 David R. Repaske,2* Gretchen L. Snyder,4 Paul Greengard,4 and Charles V. Vorhees1* Divisions of 1Developmental Biology and 2Endocrinology, Children’s Hospital Research Foundation, Cincinnati, Ohio 45229, 3Department of Biology, College of Mount St. Joseph, Cincinnati, Ohio 45233, and 4Laboratory of Molecular and Cellular Neuroscience, Rockefeller University, New York, New York 10021 Using homologous recombination, we generated mice lack- maze spatial-learning deficits. These results indicate that en- ing phosphodiesterase-mediated (PDE1B) cyclic nucleotide- hancement of cyclic nucleotide signaling by inactivation of hydrolyzing activity. PDE1B Ϫ/Ϫ mice showed exaggerated PDE1B-mediated cyclic nucleotide hydrolysis plays a signifi- hyperactivity after acute D-methamphetamine administra- cant role in dopaminergic function through the DARPP-32 and tion. Striatal slices from PDE1B Ϫ/Ϫ mice exhibited increased related transduction pathways. levels of phospho-Thr 34 DARPP-32 and phospho-Ser 845 Key words: phosphodiesterases; DARPP-32; dopamine- GluR1 after dopamine D1 receptor agonist or forskolin stimu- stimulated locomotor activity; spatial learning and memory; lation. PDE1B Ϫ/Ϫ and PDE1B ϩ/Ϫ mice demonstrated Morris Morris water maze; methamphetamine; SKF81297; forskolin Calcium/calmodulin-dependent phosphodiesterases (CaM- (CaMKII) and calcineurin and have the potential to activate PDEs) are members of one of 11 families of PDEs (Soderling et CaM-PDEs. Dopamine D1 or D2 receptor activation leads to al., 1999;Yuasa et al., 2001) and comprise the only family that acts adenylyl cyclase activation or inhibition, respectively (Traficante ϩ as a potential point of interaction between the Ca 2 and cyclic et al., 1976; Monsma et al., 1990; Cunningham and Kelley, 1993; nucleotide signaling pathways.
    [Show full text]
  • Targets and Mechanisms Validated Trials on the Horizon Targets And
    TargetsTargets and and Mechanisms Mechanisms Validated Validated TrialsTrials on on the the Horizon Horizon Research Investors Report 2011 TABLE OF CONTENTS Huntington’s Disease Research in 2011: Targets and Mechanisms Validated; Trials on the Horizon FINDING AND VALIDATING TARGETS 4 DRP1 4 Ku705 5 A CLOSER LOOK AT THE HD PROTEIN 6 HR Protein aggregates visualized 6 Form of the HD protein associated with neurodegeneration identified 7 DISCOVERING/DEVELOPING NEW DRUGS AND UNDERSTANDING THEIR MECHANISMS 10 Dantrolene appears to be neuroprotective 10 Melatonin delays onset and prolongs survival in the R6/2 Mouse 11 KMO Inhibitor developed 12 New Caspase Inhibitors identified and Optimized 14 Quinazoline derivative looks promising 15 Phosphodiesterase-10 inhibitors 16 Novel benzoxazine compounds may be neuroprotective 18 Dimethylfumarate is helpful to the YAC128 Mouse 18 IPSC Consortium creates stem cell lines 19 FINDING AND VALIDATING BIOMARKERS 21 H2AFY 21 Protein Aggregates 22 TRACK-HD 22 UNDERSTANDING THE DISEASE COURSE AND IMPROVING CLINICAL MEASURES 25 Progression of HD and MRI imaging 25 Enroll-HD 25 POTENTIAL TREATMENTS MOVING CLOSE TO CLINICAL TRIALS 27 RNAi Primate Study 27 ASOs can be made allele specific 27 Mesenchymal stem cells with BDNF 28 CLINICAL TRIALS 30 Lessons from Dimebon 30 Neurosearch -- Huntexil (ACR-16) 31 Prana Biotech Copper Chelator 31 Siena Biotech Sirt1 Inhibitor 32 THE OTHER MEMBERS OF THE RESEARCH TEAM - THE PARTICIPANTS 33 FINAL THOUGHTS 35 HD DRUG DEVELOPMENT PIPELINE CHART 36 HDSA COALITION FOR THE CURE 37 HDSA CENTERS OF EXCELLENCE 38 Cover Photo: Polarizing optical microscopy image of huntingtin peptide aggregates stained with Congo Red. Courtesy of Dr.
    [Show full text]
  • The Rational Search for PDE10A Inhibitors from Sophora Flavescens Roots Using Pharmacophore‑ and Docking‑Based Virtual Screening
    388 MOLECULAR MEDICINE REPORTS 17: 388-393, 2018 The rational search for PDE10A inhibitors from Sophora flavescens roots using pharmacophore‑ and docking‑based virtual screening HAN-TIAN FAN1, JUN-FANG GUO1, YU-XIN ZHANG2, YU-XI GU1, ZHONG-QI NING1, YAN-JIANG QIAO2 and XING WANG1,3 1School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069; 2Key Laboratory of TCM-Information Engineer of State Administration of TCM, School of Chinese Materia Medica, Beijing University of Chinese Medicine, Beijing 100102; 3Beijing Key Laboratory of Traditional Chinese Medicine (TCM) Collateral Disease Theory Research, Capital Medical University, Beijing 100069, P.R. China Received May 27, 2017; Accepted August 31, 2017 DOI: 10.3892/mmr.2017.7871 Abstract. Phosphodiesterase 10A (PDE10A) has been Introduction confirmed to be an important target for the treatment of central nervous system (CNS) disorders. The purpose of the Phosphodiesterases (PDEs) are a family of enzymes that are present study was to identify PDE10A inhibitors from herbs able to lyse phosphodiester bonds, are expressed widely, and used in traditional Chinese medicine. Pharmacophore and have demonstrable clinical significance (1,2). PDE10A has molecular docking techniques were used to virtually screen been demonstrated to be a potential target for the treatment the chemical molecule database of Sophora flavescens, a of central nervous system (CNS) disorders (3,4). Previous well-known Chinese herb that has been used for improving studies have confirmed that PDE10A inhibitors have impor- mental health and regulating the CNS. The pharmacophore tant biological activity in the treatment of psychosis (5-7). model generated recognized the common functional groups of Therefore, screening for PDE10A inhibitors is an effective known PDE10A inhibitors.
    [Show full text]
  • Phosphodiesterase (PDE)
    Phosphodiesterase (PDE) Phosphodiesterase (PDE) is any enzyme that breaks a phosphodiester bond. Usually, people speaking of phosphodiesterase are referring to cyclic nucleotide phosphodiesterases, which have great clinical significance and are described below. However, there are many other families of phosphodiesterases, including phospholipases C and D, autotaxin, sphingomyelin phosphodiesterase, DNases, RNases, and restriction endonucleases, as well as numerous less-well-characterized small-molecule phosphodiesterases. The cyclic nucleotide phosphodiesterases comprise a group of enzymes that degrade the phosphodiester bond in the second messenger molecules cAMP and cGMP. They regulate the localization, duration, and amplitude of cyclic nucleotide signaling within subcellular domains. PDEs are therefore important regulators ofsignal transduction mediated by these second messenger molecules. www.MedChemExpress.com 1 Phosphodiesterase (PDE) Inhibitors, Activators & Modulators (+)-Medioresinol Di-O-β-D-glucopyranoside (R)-(-)-Rolipram Cat. No.: HY-N8209 ((R)-Rolipram; (-)-Rolipram) Cat. No.: HY-16900A (+)-Medioresinol Di-O-β-D-glucopyranoside is a (R)-(-)-Rolipram is the R-enantiomer of Rolipram. lignan glucoside with strong inhibitory activity Rolipram is a selective inhibitor of of 3', 5'-cyclic monophosphate (cyclic AMP) phosphodiesterases PDE4 with IC50 of 3 nM, 130 nM phosphodiesterase. and 240 nM for PDE4A, PDE4B, and PDE4D, respectively. Purity: >98% Purity: 99.91% Clinical Data: No Development Reported Clinical Data: No Development Reported Size: 1 mg, 5 mg Size: 10 mM × 1 mL, 10 mg, 50 mg (R)-DNMDP (S)-(+)-Rolipram Cat. No.: HY-122751 ((+)-Rolipram; (S)-Rolipram) Cat. No.: HY-B0392 (R)-DNMDP is a potent and selective cancer cell (S)-(+)-Rolipram ((+)-Rolipram) is a cyclic cytotoxic agent. (R)-DNMDP, the R-form of DNMDP, AMP(cAMP)-specific phosphodiesterase (PDE) binds PDE3A directly.
    [Show full text]
  • (12) Patent Application Publication (10) Pub. No.: US 2003/0082511 A1 Brown Et Al
    US 20030082511A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2003/0082511 A1 Brown et al. (43) Pub. Date: May 1, 2003 (54) IDENTIFICATION OF MODULATORY Publication Classification MOLECULES USING INDUCIBLE PROMOTERS (51) Int. Cl." ............................... C12O 1/00; C12O 1/68 (52) U.S. Cl. ..................................................... 435/4; 435/6 (76) Inventors: Steven J. Brown, San Diego, CA (US); Damien J. Dunnington, San Diego, CA (US); Imran Clark, San Diego, CA (57) ABSTRACT (US) Correspondence Address: Methods for identifying an ion channel modulator, a target David B. Waller & Associates membrane receptor modulator molecule, and other modula 5677 Oberlin Drive tory molecules are disclosed, as well as cells and vectors for Suit 214 use in those methods. A polynucleotide encoding target is San Diego, CA 92121 (US) provided in a cell under control of an inducible promoter, and candidate modulatory molecules are contacted with the (21) Appl. No.: 09/965,201 cell after induction of the promoter to ascertain whether a change in a measurable physiological parameter occurs as a (22) Filed: Sep. 25, 2001 result of the candidate modulatory molecule. Patent Application Publication May 1, 2003 Sheet 1 of 8 US 2003/0082511 A1 KCNC1 cDNA F.G. 1 Patent Application Publication May 1, 2003 Sheet 2 of 8 US 2003/0082511 A1 49 - -9 G C EH H EH N t R M h so as se W M M MP N FIG.2 Patent Application Publication May 1, 2003 Sheet 3 of 8 US 2003/0082511 A1 FG. 3 Patent Application Publication May 1, 2003 Sheet 4 of 8 US 2003/0082511 A1 KCNC1 ITREXCHO KC 150 mM KC 2000000 so 100 mM induced Uninduced Steady state O 100 200 300 400 500 600 700 Time (seconds) FIG.
    [Show full text]
  • I STRUCTURE and FUNCTION of the PALMITOYLTRANSFERASE
    STRUCTURE AND FUNCTION OF THE PALMITOYLTRANSFERASE DHHC20 AND THE ACYL COA HYDROLASE MBLAC2 A Dissertation Presented to the Faculty of the Graduate School Of Cornell University In Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy By Martin Ian Paguio Malgapo December 2019 i © 2019 Martin Ian Paguio Malgapo ii STRUCTURE AND FUNCTION OF THE PALMITOYLTRANSFERASE DHHC20 AND THE ACYL COA HYDROLASE MBLAC2 Martin Ian Paguio Malgapo, Ph.D. Cornell University 2019 My graduate research has focused on the enzymology of protein S-palmitoylation, a reversible posttranslational modification catalyzed by DHHC palmitoyltransferases. When I started my thesis work, the structure of DHHC proteins was not known. I sought to purify and crystallize a DHHC protein, identifying DHHC20 as the best target. While working on this project, I came across a protein of unknown function called metallo-β-lactamase domain-containing protein 2 (MBLAC2). A proteomic screen utilizing affinity capture mass spectrometry suggested an interaction between MBLAC2 (bait) and DHHC20 (hit) in HEK-293 cells. This finding interested me initially from the perspective of finding an interactor that could help stabilize DHHC20 into forming better quality crystals as well as discovering a novel protein substrate for DHHC20. I was intrigued by MBLAC2 upon learning that this protein is predicted to be palmitoylated by multiple proteomic screens. Additionally, sequence analysis predicts MBLAC2 to have thioesterase activity. Taken together, studying a potential new thioesterase that is itself palmitoylated was deemed to be a worthwhile project. When the structure of DHHC20 was published in 2017, I decided to switch my efforts to characterizing MBLAC2.
    [Show full text]
  • Potent Lipolytic Activity of Lactoferrin in Mature Adipocytes
    Biosci. Biotechnol. Biochem., 77 (3), 566–571, 2013 Potent Lipolytic Activity of Lactoferrin in Mature Adipocytes y Tomoji ONO,1;2; Chikako FUJISAKI,1 Yasuharu ISHIHARA,1 Keiko IKOMA,1;2 Satoru MORISHITA,1;3 Michiaki MURAKOSHI,1;4 Keikichi SUGIYAMA,1;5 Hisanori KATO,3 Kazuo MIYASHITA,6 Toshihide YOSHIDA,4;7 and Hoyoku NISHINO4;5 1Research and Development Headquarters, Lion Corporation, 100 Tajima, Odawara, Kanagawa 256-0811, Japan 2Department of Supramolecular Biology, Graduate School of Nanobioscience, Yokohama City University, 3-9 Fukuura, Kanazawa-ku, Yokohama, Kanagawa 236-0004, Japan 3Food for Life, Organization for Interdisciplinary Research Projects, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan 4Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kamigyou-ku, Kyoto 602-8566, Japan 5Research Organization of Science and Engineering, Ritsumeikan University, 1-1-1 Nojihigashi, Kusatsu, Shiga 525-8577, Japan 6Department of Marine Bioresources Chemistry, Faculty of Fisheries Sciences, Hokkaido University, 3-1-1 Minatocho, Hakodate, Hokkaido 041-8611, Japan 7Kyoto City Hospital, 1-2 Higashi-takada-cho, Mibu, Nakagyou-ku, Kyoto 604-8845, Japan Received October 22, 2012; Accepted November 26, 2012; Online Publication, March 7, 2013 [doi:10.1271/bbb.120817] Lactoferrin (LF) is a multifunctional glycoprotein resistance, high blood pressure, and dyslipidemia. To found in mammalian milk. We have shown in a previous prevent progression of metabolic syndrome, lifestyle clinical study that enteric-coated bovine LF tablets habits must be improved to achieve a balance between decreased visceral fat accumulation. To address the energy intake and consumption. In addition, the use of underlying mechanism, we conducted in vitro studies specific food factors as helpful supplements is attracting and revealed the anti-adipogenic action of LF in pre- increasing attention.
    [Show full text]
  • Phosphodiesterase 4B: Master Regulator of Brain Signaling
    cells Review Phosphodiesterase 4B: Master Regulator of Brain Signaling Amy J. Tibbo and George S. Baillie * Institute of Cardiovascular and Medical Sciences, University of Glasgow, Glasgow G12 8QQ, UK; [email protected] * Correspondence: [email protected]; Tel.: +44-(0)141-330-1662 Received: 3 February 2020; Accepted: 14 May 2020; Published: 19 May 2020 Abstract: Phosphodiesterases (PDEs) are the only superfamily of enzymes that have the ability to break down cyclic nucleotides and, as such, they have a pivotal role in neurological disease and brain development. PDEs have a modular structure that allows targeting of individual isoforms to discrete brain locations and it is often the location of a PDE that shapes its cellular function. Many of the eleven different families of PDEs have been associated with specific diseases. However, we evaluate the evidence, which suggests the activity from a sub-family of the PDE4 family, namely PDE4B, underpins a range of important functions in the brain that positions the PDE4B enzymes as a therapeutic target for a diverse collection of indications, such as, schizophrenia, neuroinflammation, and cognitive function. Keywords: phosphodiesterase; cyclic-AMP; rolipram; PDE4B; neuroinflammation 1. Introduction Cyclic nucleotides are ubiquitous signaling molecules that are recognized as archetypal second messengers. Since their discovery, there has been an unprecedented drive to understand signal transduction systems that utilize them and to characterize physiological systems under their control. It is well established that both cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) signaling systems underpin critical pathways necessary for brain development and function [1–4].
    [Show full text]
  • Downregulation of Carnitine Acyl-Carnitine Translocase by Mirnas
    Page 1 of 288 Diabetes 1 Downregulation of Carnitine acyl-carnitine translocase by miRNAs 132 and 212 amplifies glucose-stimulated insulin secretion Mufaddal S. Soni1, Mary E. Rabaglia1, Sushant Bhatnagar1, Jin Shang2, Olga Ilkayeva3, Randall Mynatt4, Yun-Ping Zhou2, Eric E. Schadt6, Nancy A.Thornberry2, Deborah M. Muoio5, Mark P. Keller1 and Alan D. Attie1 From the 1Department of Biochemistry, University of Wisconsin, Madison, Wisconsin; 2Department of Metabolic Disorders-Diabetes, Merck Research Laboratories, Rahway, New Jersey; 3Sarah W. Stedman Nutrition and Metabolism Center, Duke Institute of Molecular Physiology, 5Departments of Medicine and Pharmacology and Cancer Biology, Durham, North Carolina. 4Pennington Biomedical Research Center, Louisiana State University system, Baton Rouge, Louisiana; 6Institute for Genomics and Multiscale Biology, Mount Sinai School of Medicine, New York, New York. Corresponding author Alan D. Attie, 543A Biochemistry Addition, 433 Babcock Drive, Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin, (608) 262-1372 (Ph), (608) 263-9608 (fax), [email protected]. Running Title: Fatty acyl-carnitines enhance insulin secretion Abstract word count: 163 Main text Word count: 3960 Number of tables: 0 Number of figures: 5 Diabetes Publish Ahead of Print, published online June 26, 2014 Diabetes Page 2 of 288 2 ABSTRACT We previously demonstrated that micro-RNAs 132 and 212 are differentially upregulated in response to obesity in two mouse strains that differ in their susceptibility to obesity-induced diabetes. Here we show the overexpression of micro-RNAs 132 and 212 enhances insulin secretion (IS) in response to glucose and other secretagogues including non-fuel stimuli. We determined that carnitine acyl-carnitine translocase (CACT, Slc25a20) is a direct target of these miRNAs.
    [Show full text]
  • Tracking Brain Palmitoylation Change: Predominance of Glial Change in a Mouse Model of Huntington’S Disease
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by Elsevier - Publisher Connector Chemistry & Biology Resource Tracking Brain Palmitoylation Change: Predominance of Glial Change in a Mouse Model of Huntington’s Disease Junmei Wan,1,4 Jeffrey N. Savas,2,4 Amy F. Roth,1 Shaun S. Sanders,3 Roshni R. Singaraja,3,5 Michael R. Hayden,3 John R. Yates III,2 and Nicholas G. Davis1,* 1Department of Pharmacology, Wayne State University, Detroit, MI 48201, USA 2Department of Chemical Physiology, The Scripps Research Institute, La Jolla, CA 92037, USA 3Department of Medical Genetics, Centre for Molecular Medicine and Therapeutics, Child and Family Research Institute, University of British Columbia, Vancouver, BC V5Z 4H4, Canada 4These authors contributed equally to this work 5Present address: Translational Laboratory in Genetic Medicine, Department of Medicine, National University of Singapore, and Agency for Science, Technology and Research, 8A Biomedical Grove, Singapore 138648, Singapore *Correspondence: [email protected] http://dx.doi.org/10.1016/j.chembiol.2013.09.018 SUMMARY a number of neurological disorders, notably in schizophrenia and mental retardation, as well as in Huntington’s disease (HD) Protein palmitoylation, a reversible lipid modification and Alzheimer disease (Young et al., 2012). Below, we describe of proteins, is widely used in the nervous system, ABE/SILAM, a proteomic strategy that profiles and quantifies with dysregulated palmitoylation being implicated brain palmitoylation change within
    [Show full text]
  • Potential Therapeutic Agents for Glioblastoma
    pharmaceuticals Concept Paper Methylxanthines: Potential Therapeutic Agents for Glioblastoma Daniel Pérez-Pérez 1,2 , Iannel Reyes-Vidal 2 , Elda Georgina Chávez-Cortez 2 , Julio Sotelo 2 and Roxana Magaña-Maldonado 2,* 1 PECEM, Faculty of Medicine, National Autonomous University of México, México City 04510, Mexico 2 Neuroimmunology and Neuro-oncology Unit, National Institute of Neurology and Neurosurgery, México City 14269, Mexico * Correspondence: [email protected]; Tel.: +52-55-5606-4040 Received: 5 July 2019; Accepted: 1 September 2019; Published: 7 September 2019 Abstract: Glioblastoma (GBM) is the most common and aggressive primary brain tumor. Currently, treatment is ineffective and the median overall survival is 20.9 months. The poor prognosis of GBM is a consequence of several altered signaling pathways that favor the proliferation and survival of neoplastic cells. One of these pathways is the deregulation of phosphodiesterases (PDEs). These enzymes participate in the development of GBM and may have value as therapeutic targets to treat GBM. Methylxanthines (MXTs) such as caffeine, theophylline, and theobromine are PDE inhibitors and constitute a promising therapeutic anti-cancer agent against GBM. MTXs also regulate various cell processes such as proliferation, migration, cell death, and differentiation; these processes are related to cancer progression, making MXTs potential therapeutic agents in GBM. Keywords: brain tumors; natural alkaloids; drug repositioning 1. Introduction Glioblastoma (GBM) is the most aggressive and most frequent primary malignant tumor of the central nervous system (CNS) [1]. It occurs more frequently in men and in people older than 55 years [2]. Pathological characteristics of GBM include cellular heterogeneity, angiogenesis, high proliferation rate, and increased migratory capacity [3].
    [Show full text]